Kupffer cell heterogeneity: functional properties of bone marrow derived and sessile hepatic macrophages.

A method and apparatus for automated analysis of transmitted and fluorescently labeled biological samples, wherein the apparatus automatically scans at a low magnification to acquire images which are analyzed to determine candidate cell objects of interest. Once candidate objects of interest are identified, further analysis is conducted automatically to process and collect data from samples having different staining agents.

Automated system for combining bright field and fluorescent microscopy

Defining the protein profiles of tissues and organs is critical to understanding the unique characteristics of the various cell types in the human body. In this study, we report on an anatomically comprehensive analysis of 4842 protein profiles in 48 human tissues and 45 human cell lines. A detailed analysis of over 2 million manually annotated, high-resolution, immunohistochemistry-based images showed a high fraction (>65%) of expressed proteins in most cells and tissues, with very few proteins (<2%) detected in any single cell type. Similarly, confocal microscopy in three human cell lines detected expression of more than 70% of the analyzed proteins. Despite this ubiquitous expression, hierarchical clustering analysis, based on global protein expression patterns, shows that the analyzed cells can be still subdivided into groups according to the current concepts of histology and cellular differentiation. This study suggests that tissue specificity is achieved by precise regulation of protein levels in space and time, and that different tissues in the body acquire their unique characteristics by controlling not which proteins are expressed but how much of each is produced.

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A global view of protein expression in human cells, tissues, and organs

A probe includes, a substantially L-shaped cylindrical probe body coupled through a cable to a measuring device and a temperature measuring section connected to the probe body. The temperature measuring section includes a flange portion connected to the probe body and an end portion extending from the flange portion and mounting a sensor mirror in the interior. The sensor mirror includes a cylindrical holder having a concave reflection surface in the interior, a coupling shaft extending from the cylindrical holder, a first sensor for measurement and a second sensor for correction supported in a front space of the cylindrical holder by lead wires, respectively, and a cover sheathing a front surface of the cylindrical holder. The respective lead wires supporting the first and second sensors pass the temperature measuring section and probe body and are electrically connected to the cable.

Ear-type clinical thermometer

Macrophage phagocytosis: use of fluorescence microscopy to distinguish between extracellular and intracellular bacteria.

The present invention is embodied in a method and algorithm for rapidly quantifying phagocytic functions using computer image analysis (CIA) of video light microscopic images. The method and algorithm involve sequential acquisition of bright field or phase contrast and epi-fluorescence video microscopic images of respective field, addition of the images, decision making, object referencing, morphological feature extraction, arithmetic operations, and statistical analysis. This invention provides significantly faster phagocytic functions analysis than manual microscopic examination and more detailed quantitative morphological data than flow cytometery.

System and method for quantifying macrophage phagocytosis by computer image analysis

An important new method for phagocytosis research, confocal scanning fluorescence light microscopy (CSFM), is demonstrated using fluorescent microspheres ingested by murine macrophages. CSFM, in combination with Nomarski differential interference contrast microscopy (DIC), can resolve microspheres inside cells from microspheres attached to the surface of cells. Further, combined CSFM and DIC images can quantitate phagocytosis by individual cells aggregated together. No other method offers these capabilities. A comparison of CSFM and conventional epifluorescence light microscopy (EFM) images shows that CSFM produces significantly higher-resolution images of microspheres than EFM, primarily because CSFM excludes the out-of-focus light artifacts of EFM.

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Confocal scanning fluorescence microscopy: a new method for phagocytosis research.

A new method is described for rapidly quantitating phagocytosis by adherent macrophages in culture using computer image analysis (CIA) of video light microscopic images. Ingestion of fluorescent microspheres by peritoneal murine macrophages is used to model phagocytosis. The grey levels of digital phase contrast and fluorescent microscopic images are used to quantitate the number of microspheres per cell. The method is semi-automatic, analyzes approximately 2 x 10(3) cells/hr, and simultaneously measures phagocytosis (microspheres/cell), cell area, and density (number of cells/mm2). CIA obtains the same microspheres/cell average as manual microscopic counting and an analytical precision of 5%. As expected, CIA found that the number of microspheres/cell linearly increases with increasing macrophage-microsphere co-culture time or increasing microsphere concentration until macrophages become saturated. CIA finds increased phagocytosis by interferon-gamma-treated cells and suppressed phagocytosis by cytochalasin B- or 4 degrees C-treated cells relative to controls, which demonstrates that CIA can resolve biological changes in macrophage phagocytosis. CIA also provides quantitative data on macrophage morphometry and density and found an increase in the cell area and density of INF treated macrophages. CIA provides significantly more phagocytic, morphometric, and density data than conventional manual microscopic counting methods or flow cytometric methods. The limitations, improvements, and future applications of this method are discussed.

Charles O. Odeyale

Papers

System and method for quantifying macrophage phagocytosis by computer image analysis

Confocal scanning fluorescence microscopy: a new method for phagocytosis research.

Computer image analysis method for rapid quantitation of macrophage phagocytosis.